Figure 5.

ADhbi-pro-NGF induces nuclear translocation of p75^ICD but not of the entire p75NTR in PC12 cells. PC12 cells were serum-deprived (a), serum-deprived and treated with 100 ng/ml NGF (b), or serum-deprived and treated with 25 ng/ml ADhbi-pro-NGF (c). Cells were washed, fixed, permeabilized, immunostained with anti-p75^ICD antibody (A) or with REX antibody (B) and visualized with confocal microscopy. Nuclei were stained with Hoechst. C: Apoptosis (white bars) and nuclear translocation of p75^ICD (gray bars) were quantified at 24, 48, and 72 hours. Bars represent the mean ± SD of three independent experiments. D: Western blot using anti-NRH2 antibody shows a band in PC12 total cell lysate (lane 5), which is barely detectable in total lysates from human entorhinal cortex (lanes 1 and 2) and frontal cortex (lanes 3 and 4). The arrow indicates apparent molecular weight of 24 kd. E: Epifluorescence NRH2 immunostaining image of PC12 cells treated with 25 ng/ml ADhbi-pro-NGF for 24 hours shows the absence of NRH2 nuclear translocation. Scale bars = 50 μm.